Correlation Of MRJP1 Gene Polymorphism, Gene Expression And Content In The Three Populations Of Apis Mellifera

Major royal jelly protein(MRJPs) were the part of water soluble protein, which were identified as major protein components of royal jelly,constituting about 46%-89% of the total protein content. It was found that MRJPs belong to one protein family consisting of a series of highly homologous proteins, which includes nine members. The experiments show that royal jelly has an important role in health care and biological function, which may be closely related to the MRJPs in royal jelly.This study was designed to analyze the expression pattern of MRJP1gene between different populations by Real-time PCR, which carried on the pinghu pulp bee, american Italian bee and Suwang No.1 bee. The relative content of MRJP1 was measured by HPLC to investigate the changes of MRJP1 in different populations and research the feasibility which the quality index of royal jelly was evaluated by MRJP1. The polymorphism of MRJP1 gene was analyzed by PCR-SSCP. Be based on the genotype distribution among breeds and association of MRJP1 content, the improved seed populations of high levels of MRJP1 were selected. The main results were as follows:1. The relative content of MRJP1 in 90 samples of three populations was measured by HPLC , the results showed that the relative content of MRJP1 in the Ep was 0.25%,which was significantly lower than in the Es (P<0.05)and high significantly lower than in the Em(P<0.01). The places of production of three populations belong to the region of Yangtze River Delta, and their geographic profiles such as climate, environment are consistent. And the samples were collected in the same period, the same nectar and the same age. The difference of MRJP1 relative content among different populations may be caused by internal causes of honeybee colony, and MRJP1 cannot be to a index to evaluate the the quality of royal jelly.2. The samples of three populations came from the the same period, the same nectar, the same area and the same age were selected to the expression pattern of MRJP1 gene by Real-time PCR. the results showed that, relative expression in the MRJP1 gene of Es was significantly higher than Ep and Em(P<0.05), and there were not significant difference between the relative expression of Ep and Em. The result showed that the level of production of royal jelly was not necessarily linked with the capacity of secreting MRJP1 for the individual. The correlation analysis results between the content of MRJP1 and expression of mRNA show that the correlation coefficient was 0.085, and it is no significant difference between the content of MRJP1 and expression of mRNA.3. According to the DNA sequence of MRJP1 in the GeneBank, the primers were designed to analyze the polymorphism of MRJP1 gene by PCR-SSCP. And polymorphic loci were detected in three pairs of primer. Meanwhile, 2 single nucleotide variation sites and three genotypes were detected in H5 primer. B gene is dominant gene, and the B allele frequency was 0.766,0.55,0.833, respectively. 4 single nucleotide variation sites and two genotypes were detected in W6 primer. D gene is dominant gene, and the D allele frequency was 0.867,0.967,0.900, respectively. 8 single nucleotide variation sites and three genotypes were detected in H6 primer. E gene is dominant gene, and the E allele frequency was 0.717,0.667,0.717, respectively.4. For the H5 primers, distribution and MRJP1 content of different genotypes exist differences in the three strains. MRJP1 content in AB genotype was significantly higher than AA and BB genotypes. Other genotypes were no significant differences in the strains and MRJP1 content. Therefore, AB genotype may be the advantageous genotype associated with MRJP1, and be a candidate genotype to screen the breed of high levels of MRJP1.